Cloning and Transformation of FvChi-14 Gene

Reverse transcription was used to create first strand of cDNA from 1 g of total RNA using the PrimerScriptTM-II 1st Strand cDNA Synthesis kit (TaKaRa Bio Inc. Dalian, China). The ORF sequence of the FvChi-14 (972bp) was amplified with LA Taq (TaKaRa Bio. Inc.). The PCR product was subsequently cloned into pMD20-T (TaKaRa Bio Inc.) and sequenced (FuZhou ShangYa BioInc., Fuzhou, China). The resulting sequence was cloned into the binary vector pCAMBIA1300-HA (CAMBIA company) using specific primers FvChi-14-clone-F and FvChi-14-clone-R containing BamH I and Spe I sites (Table S1). The sequence was also submitted to GenBank (Acc. NO. OQ211094). By using the freeze–thaw technique, the recombinant plasmid was transformed into Agrobacterium tumefaciens GV3101 [57 (link)], and then the recombinant was transferred into wild A. thaliana by floral dip method [58 (link)]. The solid MS medium containing 50 mg/L hygromycin was used for the screening of transgenic plants [59 (link)].